g(m1)-ganglioside and sialosylparagloboside

Unlike CNS myelin, human peripheral nerve myelin has the acidic glycosphingolipids sialosyl paragloboside (SPG), sialosyl lactosaminyl paragloboside (SLPG), and sulfated glucuronyl paragloboside (SGPG). To elucidate the pathogenesis of Guillain-Barré syndrome (GBS) and chronic inflammatory demyelinating neuropathy (CIDP), we investigated the autoantibodies to peripheral nerve molecules in patients with these diseases and compared the frequency of the autoantibodies with that of autoantibody to GM1 which is present in both the CNS and PNS. The report of Sheikh et al. (Ann. Neurol. 1995; 38: 350) that Campylobacter jejuni bears the SGPG epitope led us to study whether sera from patients with GBS subsequent to C. jejuni enteritis have anti-SGPG antibody; but, high anti-SGPG antibody titers were not found in the GBS patients from whom C. jejuni was isolated. Although the frequency of the anti-SPG, anti-SLPG and anti-SGPG antibodies were lower than that of the anti-GM1 antibody in GBS, 5 patients with demyelinating GBS had high IgG anti-SPG antibody titers. IgG anti-SPG antibody may function in the development of demyelinating GBS. We found that 6 CIDP patients had elevated IgM anti-SGPG antibody titers. Immunoelectrophoresis failed to detect IgM M-protein in 3 of the patients. IgM anti-SGPG antibody could be a diagnostic marker for a subgroup of CIDP with or without paraprotein.

Topics: Aged; Antibody Specificity; Antigens, Bacterial; Autoantibodies; Autoantigens; Autoimmune Diseases; Biomarkers; Campylobacter jejuni; Carbohydrate Sequence; Chronic Disease; Cross Reactions; Demyelinating Diseases; Epitopes; Female; G(M1) Ganglioside; Globosides; Humans; Immunoelectrophoresis; Immunoglobulin G; Immunoglobulin M; Inflammation; Lipopolysaccharides; Male; Middle Aged; Molecular Mimicry; Molecular Sequence Data; Peripheral Nerves; Polyradiculoneuropathy

The effects of various glycolipids on the activity of immunoaffinity-purified calf thymus DNA polymerase alpha were studied in vitro. Preincubation with sialic acid-containing glycolipids, such as sialosylparagloboside (SPG), GM3, GM1, and GD1a, and sulfatide (cerebroside sulfate ester, CSE) dose-dependently inhibited the activity of DNA polymerase alpha, while other glycolipids, as well as free sphingosine and ceramide did not. About 50% inhibition was achieved by preincubating the enzyme with 2.5 microM of CSE, 50 microM of SPG or GM3, and 80 microM of GM1. Inhibition was noncompetitive with both the DNA template and the substrate dTTP, as well as with the other dNTPs. Since the inhibition was largely reversed by the addition of 0.05% Nonidet P40, these glycolipids may interact with the hydrophobic region of the enzyme protein. Apparently, the sulfate moiety in CSE and the sialic acid moiety in gangliosides were essential for the inhibition since neither neutral glycolipids (i.e., glucosylceramide, galactosylceramide, lactosylceramide) nor asialo-gangliosides (GA1 and GA2) showed any inhibitory effect. Furthermore, the ceramide backbone was also found to be necessary for maximal inhibition since the inhibition was largely abolished by substituting the lipid backbone with cholesterol. Increasing the number of sialic acid moieties per molecule further enhanced the inhibition, while elongating the sugar chain diminished it. It was clearly shown that the N-acetyl residue of the sialic acid moiety is particularly essential for inhibition by both SPG and GM3 because the loss of this residue or substitution with a glycolyl residue completely negated their inhibitory effect on DNA polymerase alpha activity.

Topics: Animals; Carbohydrate Sequence; Cattle; DNA Polymerase II; G(M1) Ganglioside; G(M3) Ganglioside; Globosides; Glycolipids; Kinetics; Molecular Sequence Data; N-Acetylneuraminic Acid; Sialic Acids; Sulfates; Thymus Gland

Liposomes as defined model membranes were used to quantitatively study the effects of specific sialic acid containing glycolipids on activation of the alternative pathway of human C. Liposomes containing dimyristoylphosphatidylethanolamine, cholesterol, and cerebrosides at molar ratios of 1.0/0.75/0.33 activated the alternative pathway in human serum treated with MgEGTA. Activation was measured by C3 conversion and the deposition of total C3 and functional C3b on the liposome surface. The monosialoganglioside GM1, when incorporated into the activating liposome membrane at molar ratios between 10(-5) and 10(-2), inhibited activation in a dose-dependent manner. Sialosylparagloboside also inhibited activation in human serum, and inhibition was completely reversed after neuraminidase treatment. The degree of inhibition by GM1 correlated with the relative amount of GM1 exposed on the liposome surface. Sialic acid did not directly inhibit the binding of C3b when liposomes containing gangliosides were incubated with the purified components C3, B, D, and P. GM1 did inhibit activation when liposomes were incubated with a mixture of purified C3, B, D, P, H, and I. Binding assays with radiolabeled H showed increased binding of H to liposome-bound C3b in the presence of GM1. These results establish the ability of sialic acid on glycolipids to promote H binding to C3b and thereby regulate alternative pathway activation on a defined lipid membrane.

Topics: Binding, Competitive; Complement Activation; Complement C3; Complement C3b Inactivator Proteins; Complement Factor B; Complement Pathway, Alternative; G(M1) Ganglioside; Gangliosides; Globosides; Humans; Lipid Bilayers; Liposomes; Models, Biological; Neuraminidase

The turnover rate of GM1 ganglioside in myelin was examined reveal age-related alterations in the metabolic activity. Three different age groups of mice were given deuterium oxide, and myelin was prepared from cerebella at intervals of two weeks. GM1 was isolated from the total myelin gangliosides by high performance liquid chromatography. Deuterated sugar moieties of GM1 were determined by chemical ionization-mass spectrometry which provided prominent quasimolecular ions. This method made it possible to determine separately the incorporation of deuterium into internal and external galactoses as well as other components. The incorporation rate of GM1 into myelin was clearly shown to be decreased with advancing age. Lower incorporation of newly synthesized sialic acid into GM1 than that of other sugars may indicate reutilization of sialic acid at about 50%. The possibility of modification of the myelin metabolism by exogenous factors was examined by monitoring the incorporation rate of GM1 in animals treated with chemical agents. It was revealed that thyroxine enhanced the incorporation of GM1 into adult brain myelin, whereas propylthiouracil reduced the incorporation. Other chemicals, estradiol, S-adenosylmethionine and LM1 ganglioside, showed only minor effects on the myelin turnover.

Topics: Aging; Animals; Brain; Ethinyl Estradiol; Female; G(M1) Ganglioside; Gangliosides; Globosides; Mice; Mice, Inbred C57BL; Myelin Sheath; Propylthiouracil; S-Adenosylmethionine; Thyroxine